Team:Sheffield/Proof
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We have designed a device () using the promoter, adding an RBS site to it, and coupling it to a GFP with a ryhB binding sequence.
In a bacterial infection, lipocalin is produced by the human or animal organism. Lipocalin prevents bacteria from uptaking iron. We use GFP expression as a means to determine and quantify the presence of bacterial infection, by utilising the ryhB pathway.
In the presence of lipocalin (thus, absence of intracellular iron), ryhB can freely degrade GFP mRNA in bacteria. In the absence of lipocalin (thus, presence of intracellular iron), the Fur regulator represses ryhB, and GFP is produced.
We have used fluorometry to experimentally validate that our engineered bacteria change their levels of GFP expression after addition of a defined amount of iron.
